Methods for cancer therapy

ABSTRACT

The present invention relates to methods for treating patients with cancer, including patients with hematological malignancy, wherein the method comprises administering to the patient a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein R1 and R2 are as defined herein.

FIELD OF THE INVENTION

The present invention relates to methods for treating patients withcancer, including patients with hematological malignancy.

BACKGROUND OF THE INVENTION

Cancer has a major impact on society across the world. Cancer is thesecond most common cause after cardiovascular disease responsible forhuman death. The National Cancer Institute estimates that in 2015,approximately 1,658,370 new cases of cancer will be diagnosed in theUnited States and 589,430 people will die from the disease.

Chronic myeloid leukemia (CML) is a type of cancer that starts incertain blood forming cells of the bone marrow. CML cells contain anabnormal gene, BCR-ABL, that isn't found in normal cells. This genemakes a protein, BCR-ABL, which causes CML cells to grow and reproduceout of control. BCR-ABL is a type of protein known as a tyrosine kinase.Drugs known as tyrosine kinase inhibitors (TKIs) that target BCR-ABL arethe standard treatment for CML.

Imatinib (Gleevec®) is the first drug to specifically target the BCR-ABLtyrosine kinase protein for treating CML. However, emerging acquiredresistance to imatinib has become a major challenge for clinicalmanagement of CML. More than 100 resistance-related BCR-ABL mutants havebeen identified in the clinic, among which the “gatekeeper” T315I ismost common mutation, as it accounts for approximately 15-20% of allclinically acquired mutants. Ren et al., J. Med Chem. 2013, 56, 879-894.

Great efforts have been devoted to identifying second generation ofBCR-ABL inhibitors to overcome imatinib resistance. Nonetheless, thesecond-generation inhibitors are not capable of inhibiting the mostrefractory BCR-ABL^(T315I) mutant. BCR-ABL^(T315I) induced drugresistance remains an unmet clinical challenge for CML treatment.Accordingly, there is a continuing need for new and more effectivetreatment. The methods of the present invention present cancer patientswith new options.

SUMMARY OF THE INVENTION

The present invention relates methods for treating cancer in a patient,comprising administering to the patient a therapeutically effectiveamount of a compound of formula (I):

or a pharmaceutically acceptable salt thereof, wherein R₁ is hydrogen,C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₁₋₄ alkyloxy, or phenyl; and R₂ ishydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or halogen.

In certain embodiments, the cancer is hematological malignancy.

In certain embodiments, the hematological malignancy is leukemia,including chronic myelogenous leukemia.

In certain embodiments, the method is in the treatment of the patientwith chronic myeloid leukemia resistant to current tyrosine kinaseinhibitor therapies.

In certain embodiments, the patient with chronic myeloid leukemiaresistant to the current tyrosine kinase inhibitor therapies is causedby BCR-ABL mutations.

In certain embodiments, BCR-ABL mutation is T315I, E255K/V, G250E,H396P, M351T, Q252H, Y253F/H, or BCR-ABL^(WT) mutations.

In certain embodiments, BCR-ABL mutation is T315I mutation.

In certain embodiments, the compound of formula (I), or pharmaceuticallyacceptable salt thereof is administered orally to the patients in needsuch treatment.

In certain embodiments, the compound of formula (I), or pharmaceuticallyacceptable salt thereof is administered once every other day (QOD)during the 28-day treatment cycle.

In one embodiment, the compound of formula (I) is a compound of formula(I-A):

or a pharmaceutically acceptable salt thereof.

In certain embodiments, the compound of formula (I) or formula (I-A) isadministered once every other day in an amount of about 1 mg, about 2mg, about 4 mg, or about 8 mg.

In certain embodiments, the compound of formula (I) or formula (I-A) isadministered once every other day in an amount of about 12 mg or about20 mg.

In certain embodiments, the compound of formula (I) or formula (I-A) isadministered once every other day in an amount of about 30 mg, about 40mg, or about 45 mg.

In certain embodiments, the compound of formula (I) or formula (I-A) isadministered once every other day in an amount of about 50 mg or about60 mg.

In certain embodiments, the present invention relates to a method ofinhibiting BCR-ABL mutants, comprising contacting a compound of formula(I) or a salt thereof with BCR-ABL mutants, wherein the BCR-ABL mutantsis T315I, E255K/V, G250E, H396P, M351T, Q252H, Y253F/H, or BCR-ABL^(WT).

In certain embodiments, the present invention relates to a method ofinhibiting BCR-ABL mutants, comprising contacting a compound of formula(I) or a salt thereof with BCR-ABL mutants selected from T315I.

In certain embodiments, the present invention provides a medicament orpharmaceutical composition comprising the compound of formula (I) orformula (I-A) or pharmaceutically acceptable salt thereof for treathematological malignancy, including chronic myelogenous leukemia.

In certain embodiments, the present invention relates to us of acompound of formula (I) or (I-A), or pharmaceutically acceptable saltthereof in the manufacture of medicament for the treatment ofhematological malignancy, including chronic myelogenous leukemia.

BRIEF DESCRIPTION OF FIGURES

FIGS. 1A and 1B illustrate efficacy (CHR n %) of the compound of formula(I-A) in a phase 1 study.

FIGS. 2A and 2B illustrate efficacy (MCyR n %) of the compound offormula (I-A) in a phase 1 study.

FIGS. 3A, 3B, and 3C illustrate efficacy (MCyR n %) by dose (CP) of thecompound of formula (I-A) in a phase 1 study.

FIGS. 4A and 4B illustrate MMR (MCyR n %) of the compound of formula(I-A) in a phase 1 study.

FIGS. 5A and 5B illustrate plasma concentration-time profiles of thecompound of formula (I-A) in a phase 1 study.

FIGS. 6A and 6B illustrate the efficacy of the compound of formula (I-A)in a phase 1 study.

FIG. 7A illustrates the response rate and the depth of response of thecompound of formula (I-A) in CML-CP patients, Example 2. FIG. 7Billustrates the progression free survival (PFS) rate of the compound offormula (I-A) in CML patients.

FIG. 8 illustrates the effect of the compound of formula (I-A) onsurvival of Ba/F3 tumor bearing mice expressing the BCR-ABL^(T315I).

DETAILED DESCRIPTION OF THE INVENTION

All published documents cited herein are hereby incorporated herein byreference in their entirety.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs.

The term “about” is used herein to mean approximately, in the region of,roughly, or around. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth. In general, the term“about” is used herein to modify a numerical value above and below thestated value by a variance of 10%.

The term “comprises” refers to “includes, but is not limited to.”

As used herein, the terms “treatment,” “treat,” and “treating” refer toreversing, alleviating, delaying the onset of, or inhibiting theprogress of a disease or disorder, or one or more symptoms thereof,including but not limited to therapeutic benefit. In some embodiments,treatment is administered after one or more symptoms have developed. Insome embodiments, treatment may be administered in the absence ofsymptoms. For example, treatment may be administered to a subject priorto the onset of symptoms (e.g., in light of a history of symptoms and/orin light of genetic or other susceptibility factors). Treatment may alsobe continued after symptoms have resolved, for example to prevent ordelay their recurrence.

Therapeutic benefit includes eradication and/or amelioration of theunderlying disorder being treated such as cancer; it also includes theeradication and/or amelioration of one or more of the symptomsassociated with the underlying disorder such that an improvement isobserved in the subject, notwithstanding that the subject may still beafflicted with the underlying disorder. In some embodiments, “treatment”or “treating” includes one or more of the following: (a) inhibiting thedisorder (for example, decreasing one or more symptoms resulting fromthe disorder, and/or diminishing the extent of the disorder); (b)slowing or arresting the development of one or more symptoms associatedwith the disorder (for example, stabilizing the disorder and/or delayingthe worsening or progression of the disorder); and/or (c) relieving thedisorder (for example, causing the regression of clinical symptoms,ameliorating the disorder, delaying the progression of the disorder,and/or increasing quality of life.)

As used herein, “administering” or “administration” of the compound offormula (I) or formula (I-A) or a pharmaceutically acceptable saltthereof encompasses the delivery to a patient a compound or apharmaceutically acceptable salt thereof, or a prodrug or otherpharmaceutically acceptable derivative thereof, using any suitableformulation or route of administration, e.g., as described herein.

As used herein, the term “therapeutically effective amount” or“effective amount” refers to an amount that is effective to elicit thedesired biological or medical response, including the amount of acompound that, when administered to a subject for treating a disorder,is sufficient to effect such treatment of the disorder. The effectiveamount will vary depending on the disorder, and its severity, and theage, weight, etc. of the subject to be treated. The effective amount maybe in one or more doses (for example, a single dose or multiple dosesmay be required to achieve the desired treatment endpoint). An effectiveamount may be considered to be given in an effective amount if, inconjunction with one or more other agents, a desirable or beneficialresult may be or is achieved. Suitable doses of any co-administeredcompounds may optionally be lowered due to the combined action, additiveor synergistic, of the compound.

As used herein, “delaying” development of a disorder mean to defer,hinder, slow, stabilize, and/or postpone development of the disorder.Delay can be of varying lengths of time, depending on the history of thedisease and/or the individual being treated.

As used herein, “patient” to which administration is contemplatedincludes, but is not limited to, humans (i.e., a male or female of anyage group, e.g., a pediatric subject (e.g., infant, child, adolescent)or adult subject (e.g., young adult, middle-aged adult or senior adult))and/or other primates (e.g., cynomolgus monkeys, rhesus monkeys).

As used herein, “pharmaceutically acceptable” or “physiologicallyacceptable” refer to compounds, salts, compositions, dosage forms andother materials which are useful in preparing a pharmaceuticalcomposition that is suitable for veterinary or human pharmaceutical use.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge etal., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66, 1-19. Pharmaceutically acceptablesalts of Compound 1 include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate,propionate, stearate, succinate, sulfate, tartrate, thiocyanate,p-toluenesulfonate, undecanoate, valerate salts, and the like. Althoughpharmaceutically acceptable counter ions will be preferred for preparingpharmaceutical formulations, other anions are quite acceptable assynthetic intermediates. Thus it may be pharmaceutically undesirableanions, such as iodide, oxalate, trifluoromethanesulfonate and the like,when such salts are chemical intermediates.

As used herein, alkyl refers to methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl, or tert-butyl. Alkyl groups can besubstituted or unsubstituted.

As used herein, cycloalkyl refers to cyclopropyl, cyclobutyl,cyclopentyl or cyclohexyl. Cycloalkyl groups can be substituted orunsubstituted.

As used herein alkoxy refers to methoxy, ethoxy, propoxy, isopropoxybutoxy, isobutoxy, sec-butoxy, or tert-butoxy. Alkoxy groups can besubstituted or unsubstituted.

As used herein, halogen refers to fluorine, chlorine, bromine or iodine.

As used herein, compound of formula (I) has the following structure:

wherein R₁ is hydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₁₋₄ alkyloxy, orphenyl; and R₂ is hydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or halogen.

As used herein, the compound of formula (I-A) has the followingstructure:

The chemical name for the compound of (I-A) is3-(2-(1H-pyrazolo[3,4-b]pyridin-5-yl)ethynyl)-4-methyl-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-benzamide.

As used herein, the compounds of formula (I) or (I-A) include anytautomer forms. As a non-limiting example, tautomerization may occur inthe pyrazole and pyrimidine groups.

The compounds of formula (I) or formula (I-A) or a pharmaceuticallyacceptable salt thereof can be obtained according to the productionmethods described in U.S. Pat. No. 8,846,671 B2, issued Sep. 30, 2014,which is incorporated herein by reference in its entirety and for allpurposes, or a method analogous thereto.

The compounds of formula (I) or (I-A) are novel, selective potentinhibitors against a broad spectrum of BCR-ABL mutations, includingT315I, E255K/V, G250E, H396P, M351T, Q252H, Y253F/H, or BCR-ABL^(WT).

The compounds of formula (I) or formula (I-A) or a pharmaceuticallyacceptable salt thereof are also potent inhibitors against other kinasesincluding KIT, BRAF, DDR1, PDGFR, FGFR, FLT3, RET, SRC, TIE1, and TIE2.

Also provided herein are pharmaceutical compositions and dosage forms,comprising compounds of formula (I) or formula (I-A) or apharmaceutically acceptable salt thereof, and one or morepharmaceutically acceptable excipients. Compositions and dosage formsprovided herein may further comprise one or more additional activeingredients. Compounds of formula (I) or formula (I-A) or apharmaceutically acceptable salt thereof may be administered as part ofa pharmaceutical composition as described.

In some embodiment, provided is a method for hematological malignancy ina patient, comprising administering to the patient a therapeuticallyeffective amount of a compound of formula (I):

or a pharmaceutically acceptable salt thereof, wherein R₁ is hydrogen,C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₁₋₄ alkyloxy, or phenyl; and R₂ ishydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or halogen.

In certain embodiments, the compound of formula (I) is a compound offormula (I-A):

or a pharmaceutically acceptable salt thereof.

In certain embodiments, the hematological malignancy is leukemia,including chronic myelogenous leukemia.

In certain embodiments, the method is in the treatment of the patientwith chronic myeloid leukemia resistant to current tyrosine kinaseinhibitor therapies, wherein resistant to the current tyrosine kinaseinhibitor therapies is caused by BCR-ABL mutations.

Examples of the current tyrosine kinase inhibitors include, but notlimit to, imatinib, dasatinib, nilotinib, bosutinib, ponatinib, orbafetinib,

In certain embodiments, BCR-ABL mutation is T315I, E255K/V, G250E,H396P, M351T, Q252H, Y253F/H, or BCR-ABL^(WT) mutations.

In certain embodiments, BCR-ABL mutation is T315I mutation.

In a more preferred embodiment of the invention, the method of theinvention relates to a method for treating a hematological malignancyresistant to Ponatinib.

Ponatinib is a third-generation inhibitor of BCR-ABL for the treatmentof Chronic Myelogenous Leukemia (CML) carrying the T315I mutation,Ph+ALL (Philadelphia chromosome positive ALL), and CML and Ph+ALL thatare not responsive to other Tyrosine Kinase Inhibitors (TKIs). AlthoughPonatinib is clinically active against most BCR-ABL single mutations, apart of patients still do not respond consistently to Ponatinib (see:Cortes J E et al. A phase 2 trial of ponatinib in Philadelphiachromosome-positive leukemias. N Engl J Med. 2013; 369(19):1783-96).Studies have shown that the compound mutation of BCR-ABL may be involvedin clinical drug resistance of CML and Ph+ALL to Ponatinib. For example,in Ph+ALL, the E255V/T315I double mutation can produce 20-foldresistance compared to the T315I single mutation, and other compoundmutations such as Q252H/T315I, T315I/M351I and T315I/F359V are also lesssensitive to Ponatinib (see: Zabriskie M S, et al. BCR-ABL1 compoundmutations combining key kinase domain positions confer clinicalresistance to ponatinib in Ph chromosome-positive leukemia. Cancer Cell.2014; 26(3):428-42). In CML, patients containing both T315I and othermutations are more resistant than patients with single mutation T315I(see: Parker W T et al. The impact of multiple low-level BCR-ABL1mutations on response to ponatinib. Blood. 2016; 127(15): 1870-80).

In a preferred embodiment of the invention, the inhibitory effect of thecompound of formula (I) and Ponatinib on BCR-ABL complex mutant cellproliferation is confirmed by constructing a stably transfected cellline with BCR-ABL complex mutation, and a potential therapeutic approachto overcome Ponatinib resistance is provided.

The invention proves that the compound of the formula (I-A) has betterantiproliferative effect than Ponatinib on Ba/F3 cells with complexmutations of BCR-ABL^(E255V/T315I), BCR-ABL^(Y253H/E255V),BCR-ABL^(T315M), BCR-ABL^(Y253H/T315I), BCR-ABL^(Y253H/F359V) andBCR-ABL^(T315I/F317L). The results suggest that the compound of formula(I-A) is a potential candidate drug for overcoming the resistance ofPonatinib caused by BCR-ABL complex mutation.

In certain embodiments, the compound of formula (I), or pharmaceuticallyacceptable salt thereof is administered orally to the patients in needsuch treatment.

In certain embodiments, the compound of formula (I), or pharmaceuticallyacceptable salt thereof is administered once every one, two, or threedays during the treatment cycle. The said treatment cycle may be 20-40days, preferably 25-35 days, more preferably 28-day treatment cycle.

In certain embodiments, the compound of formula (I) or formula (I-A) isadministered every day, or once every other day (QOD), or once everythree days, particularly once every other day. The amount ofadministration is from 0.5 mg to 100 mg, preferably from 1 mg to 80 mg,more preferably from 1 mg to 60 mg. In the most preferable embodiments,it is in an amount of about 1 mg, 2 mg, 4 mg, 6 mg, 8 mg, 10 mg, 12 mg,14 mg, 16 mg, 18 mg, 20 mg, 22 mg, 24 mg, 26 mg, 28 mg, 30 mg, 32 mg, 34mg, 36 mg, 38 mg, 40 mg, 42 mg, 44 mg, 46 mg, 48 mg, 50 mg, 52 mg, 54mg, 56 mg, 58 mg, or 60 mg.

In certain embodiments, the compound of formula (I) or formula (I-A) isadministered once every other day in an amount of about 30 mg, about 40mg, or about 45 mg.

In certain embodiments, the compound of formula (I) or formula (I-A) isadministered once every other day in an amount of about 50 mg or about60 mg.

In certain embodiments, the compound of formula (I) or formula (I-A) isformulated into a dosage unit to be administered every day, or onceevery other day (QOD), or once every three days, particularly once everyother day. The amount of the dosage unit is from 0.5 mg to 100 mg,preferably from 1 mg to 80 mg, more preferably from 1 mg to 60 mg.

In certain embodiments, the present invention relates to a method ofinhibiting BCR-ABL mutants, comprising contacting a compound of formula(I) or a salt thereof with BCR-ABL mutants, wherein the BCR-ABL mutantsis T315I, E255K/V, G250E, H396P, M351T, Q252H, Y253F/H, or BCR-ABL^(WT).

In certain embodiments, the present invention relates to a method ofinhibiting BCR-ABL mutants, comprising contacting a compound of formula(I) or a salt thereof with BCR-ABL mutants selected from T315I.

In certain embodiments, the inhibition is in vitro or in vivo.

In certain embodiments, the inhibition is in a patient with chronicmyeloid leukemia resistant to current tyrosine kinase inhibitortherapies.

In certain embodiments, the present invention provides a medicament orpharmaceutical composition comprising the compound of formula (I) orformula (I-A) or pharmaceutically acceptable salt thereof for treathematological malignancy, including chronic myelogenous leukemia.

In certain embodiments, the present invention relates to the use of acompound of formula (I) or (I-A), or pharmaceutically acceptable saltthereof in the manufacture of medicament for the treatment ofhematological malignancy, including chronic myelogenous leukemia.

In certain embodiments, the compound of formula (I) or (I-A) is in asolid dosage form.

In certain embodiments, the cancer is newly diagnosed.

In certain embodiments, the cancer is relapsed.

In certain embodiments, the cancer is refractory.

The present disclosure describes various embodiments. A person ofordinary skill in the art reviewing the disclosure will readilyrecognize that various embodiments can be combined in any variation. Forexample, embodiments of the disclosure include treatment of variousdisorders, patient populations, administrations of dosage forms, atvarious dosages, minimization of various adverse events, andimprovements in various efficacy measures, etc. Any combinations ofvarious embodiments are within the scope of the disclosure.

As used herein, the term “survival” refers to the patient remainingalive, and includes progression-free survival (PFS) and overall survival(OS). Survival can be estimated by the Kaplan-Meier method, and anydifferences in survival are computed using the stratified log-rank test.

As used herein, the term “progression-free survival (PFS)” refers to thetime from treatment (or randomization) to first disease progression ordeath. For example it is the time that the patient remains alive,without return of the cancer (e.g., for a defined period of time such asabout one month, two months, three months, three and a half months, fourmonths, five months, six months, seven months, eight months, ninemonths, about one year, about two years, about three years, about fiveyears, about 10 years, about 15 years, about 20 years, about 25 years,etc.) from initiation of treatment or from initial diagnosis.Progression-free survival can be assessed by Response EvaluationCriteria in Solid Tumors (RECIST).

The term “overall survival” refers to the patient remaining alive for adefined period of time (such as about one year, about two years, aboutthree years, about four years, about five years, about 10 years, about15 years, about 20 years, about 25 years, etc.) from initiation oftreatment or from initial diagnosis.

Non-limiting examples of hematologic malignancies also includeamyloidosis, acute myeloid leukemia (AML); chronic myelogenous leukemia(CML) including accelerated CML and CML blast phase (CML-BP); acutelymphoblastic leukemia (ALL); chronic lymphocytic leukemia (CLL);Hodgkin's disease (HD); non-Hodgkin's lymphoma (NHL), includingfollicular lymphoma and mantle cell lymphoma; B-cell lymphoma; T-celllymphoma; multiple myeloma (MM); Waldenstrom's macroglobulinemia;myelodysplastic syndromes (MDS), refractory anemia (RA), refractoryanemia with ringed siderblasts (RARS), refractory anemia with excessblasts (RAEB), and RAEB in transformation (RAEB-T); andmyeloproliferative syndromes.

For cancer therapy, efficacy may be measured by assessing the durationof survival, duration of progression-free survival (PFS), the responserates (RR) to treatments, duration of response, and/or quality of life.

The term “pharmaceutically acceptable carrier” is used herein to referto a material that is compatible with a recipient subject, preferably amammal, more preferably a human, and is suitable for delivering anactive agent to the target site without terminating the activity of theagent. The toxicity or adverse effects, if any, associated with thecarrier preferably are commensurate with a reasonable risk/benefit ratiofor the intended use of the active agent.

The pharmaceutical compositions of this disclosure can be manufacturedby methods well known in the art such as conventional granulating,mixing, dissolving, encapsulating, lyophilizing, or emulsifyingprocesses, among others. Compositions may be produced in various forms,including granules, precipitates, particulates, or powders.

The term “orally” refers to administering a composition that is intendedto be ingested. Examples of oral forms include, but are not limited to,tablets, pills, capsules, powders, granules, solutions or suspensions,and drops. Such forms may be swallowed whole or may be in chewable form.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activeingredient is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid; b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia; c) humectants such as glycerol; d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate; e) solutionretarding agents such as paraffin; f) absorption accelerators such asquaternary ammonium compounds; g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate; h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents such as phosphates orcarbonates.

Solid compositions may also be employed as fillers in soft andhard-filled gelatin capsules using such excipients as lactose or milksugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition such that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

In solid dosage forms the active ingredients may be mixed with at leastone inert diluent such as sucrose, lactose or starch. Such dosage formsmay also comprise, as is normal practice, additional substances otherthan inert diluents, e.g., tableting lubricants and other tableting aidssuch a magnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents.

The active ingredients can also be in micro-encapsulated form with oneor more excipients as noted above.

EXAMPLES Example 1

A single-agent, open-label dose escalation and dose expansion Phase Istudy to assess the safety, preliminary efficacy, pharmacokinetic (PK)and pharmacodynamic (PD) properties of orally administered the compoundof formula (I-A) in the TKI-resistant patients with chronic phase (CP)or accelerated phase (AP) CML.

Methods: the compound of formula (I-A) was administered orally onceevery other day (QOD) in 28-days cycles at 11 dose cohorts ranging from1 mg to 60 mg. The eligible patients received treatments until diseaseprogression or intolerable toxicities. The primary efficacy endpoint inthe CML AP and CP patients, was complete hematological response (CHR)and major cytogenetic response (MCyR) respectively, MCyR includespartial cytogenetic response (PCyR) and complete cytogenetic response(CCyR). Blood samples were collected at various time points on Day 1-2and Day 27-28 during cycle 1 for PK analyses. BCR-ABL inhibition wasevaluated using tyrosine phosphorylation of CRKL and STAT5 in peripheralblood mononuclear cell (PBMCs) collected from the patients before and 4,8, 24 and 48 hours post the compound of formula (I-A) treatments on Day1, 15 and 27 during cycle 1.

Dose Escalation:

-   -   3 to 6 subjects in each dose group (traditional 3+3 dose        increasing method)    -   Up to 15 subjects in the MTD dose group (safety and PK        characteristics)    -   Dose extension studies:    -   Up to 60 subjects (CP/CML-AP)    -   Extended dose selection is based on initial safety and efficacy

Study Endpoints

-   -   Primary endpoint    -   To determine the safety and RP2D (recommended phase 2 dose) of        the compound of formula (I-A) in patients with        resistant/refractory CML    -   Secondary endpoints    -   To examined the safety of the compound of formula (I-A) in        resistant/refractory CML patients.    -   To evaluate the pharmacokinetic characteristics of the compound        of formula (I-A)    -   To evaluate the efficacy of the compound of formula (I-A) in        resistant/refractory CML patients.

In certain embodiments, patients who meet the following criteria mayreceive the treatments:

-   -   Patients >18 years in CP or AP.    -   TKI resistance.    -   ECOG≥2.    -   Minimum life expectancy of ≥3 months.    -   Patients with adequate organ function.    -   Heart function: Left ventricular ejection fraction (LVEF) >50%.    -   EKG QTc interval: male ≤450 ms, female ≤470 ms.    -   Agree to use effective form of contraception (as applicable).    -   Ability to comply with study procedures, in the Investigator's        opinion.

In certain embodiments, patients who has the following criteria mayexclude from the treatments:

-   -   The patients who received cytotoxic chemotherapy, or any other        radiotherapy within 28 days; or interferon, cytarabine within 14        days; or other TKIs within 14 days; or any adverse events (AEs)        not recovered to CTCAE grade 0-1 due to any other treatments        (except alopecia).    -   Patients who are currently receiving treatment with a medication        that has the potential to interact with the compound of formula        (I-A).    -   Patients who had been treated with ponatinib, or the compound of        formula (I-A) like drugs.    -   Impairment of gastrointestinal (GI) function or GI disease that        may significantly alter absorption of study drugs.    -   Patients with cardiovascular diseases, including HBP (>140/90        mmHg); or receiving drugs that can cause prolonged QT interval.    -   Mean pulmonary artery pressure >25 mmHg.    -   Having experience of serious cardiovascular AEs during previous        TKI treatment.    -   Patients having history of HSCT.    -   Patients having abnormal coagulation function or having a        significant bleeding disorder unrelated to CML.    -   Patients who have a major surgery within 4 weeks prior to study        entry.    -   Patients who have a history of bleeding disease unrelated to        CML.    -   Patients who require immunosuppressive therapy other than short        time of steroid.    -   Cytologically confirmed central nervous system (CNS) involvement        (if asymptomatic, spinal fluid examination is not necessary        prior to first treatment).    -   Patients with the medical history of clinically significant        primary malignancy concurrently clinically significant primary        malignancy.    -   Have ongoing or active infection, including HIV+, Hepatitis A,        B, or C.    -   Known allergy to any components in the study drug.    -   Pregnant or lactating.

Patients' Characteristics

Total CP AP Age, median 40 (20-64) 39 (20-64) 40 (20-64) (range) Male, n(%) 70 (70%) 58 (67%) 12 (86%) ECOG, n (%)  0 84 (84%) 71 (83%) 13 (93%) 1 16 (16%) 15 (17%)  1 (7%) Interval Dx to  6 (0.3-15.2)  5 (0.6-15.2) 8 (0.3-14.7) the compound of formula (I-A), median (range) PriorTKI-therapy, n (%)  1 17 (17%) 12 (14%)  5 (36%)  2 51 (51%) 47 (55%)  4(29%) ≥3 32 (32%) 27 (31%)  5 (36%) TKI type Imatinib or 12 (12%) 12(14%)  1 (36%) Nilotinib Imatinib + 32 (32%) 30 (35%)  2 (14%) DasatinibImatinib + 17 (17%) 15 (17%)  2 (14%) Nilotinib Imatinib +  5 (5%) 27(31%)  5 (36%) Dasatinib + Nilotinib

BCR-ABL Mutation Status at Baseline

Total CP AP No mutation,  23(23%) 22 (26%)  1 (7%) n (%) Mutation(s) n(%)  1 17 (17%) 12(14%)  5(36%)  2 51 (51%) 47(55%)  4(29%) ≥3 14 (14%)10(12%)  4(29%) Type T315I 13 (63%) 52 (60%) 11 (19%) F317L 11 (11%)  9(10%)  2 (14%) E255K  7 (7%)  5 (6%)  2 (14%) 359I/V  3 (3%)  2 (2%)  1(7%) 244V  4 (4%)  3 (3%)  1 (7%) G250E  3 (3%)  3 (3%)  0 (%) E459K  2(2%)  2 (2%)  0 (%) Y253F/H  2 (2%)  2 (2%)  0 (%) Others: 10 (10%) 10(38)  0 (%)

Patent Disposition

Patients-n CP AP Total Continuing on 85(99%) 10(71%) 95(95%) treatment,n(%) Not continuing on treatment, but continuing in study, n (%)PD/Failure  0  4(29%)  4(4%) Others  1(1%)  0  1(1%)

Treatment Related Adverse Events

Treatment related Adverse events in 10% of patients Any grade Grade 3-4SAE n (%) n (%) n (%) leukocytosis  9 (9.0%)  8 (8.0%) 0 Leukopenia 17(17%) 15 (15%) 0 Neutropenia  2 (2.0%)  0 0 Thrombocytopenia 46 (46%) 45(45%) 5 (5.0%) Anaemia 10 (10%)  9 (9.0%) 2 (2.0%) Proteinuria 11 (11%) 1 (1.0%) 0 Hypertriglyceridaemia 51 (51%)  4 (4.0%) 0 Skinhyperpigmentation 35 (35%)  0 0 Alanine aminotransferase 29 (29%)  2(2.0%) 0 increased Aspartate aminotransferase 31 (31%)  2 (2.0%) 0inceased Hypocalcaemia 31 (31%)  0 0 Blood bilirubin increased 28 (28%) 1 (1.0%) 0 Blood creatine phosphokinase 19 (19%)  1 (1.0%) 0 increased

On Treatment Period

Dose Number On-treatment duration cohort of patients median range  1 mg3 12.6 12.3-16.5  2 mg 3 9.6  2.0-15.3  4 mg 3 8.5  8.4-9.3  8 mg 3 10.4 7.3-13.3 12 mg 3 11.4 11.4-12.9 20 mg 4 10.9  8.4-14.0 30 mg 37 7.4 0.0-11.4 40 mg 37 3.7  1.2-7.0 45 mg 3 11.3 11.1-11.3 50 mg 24 5.2 0.7-7.2 60 mg 3 9.4  9.4-9.7

Treatment Related AEs Leading to Dose Interruption/Reduction by Dose

40 mg 45 mg 50 mg 60 mg 20 mg(n = 4) 30 mg(n = 37) (n = 37) (n = 3) (n =24) (n = 3) AE n(%) n(%) n(%) n(%) n(%) n(%) Thrombocytopenia 1(25%) 12(32%) 11 (30%) 2(67%) 8 (33%) 2(67%) Hypocalcemia 0 2 (5.4%) 0 0 1(4.2%) 0 AST increased 0 0 1 (2.7%) 0 1 (4.2%) 0 ALT increased 0 0 1(2.7%) 0 1 (4.2%) 0 Lipase increased 0 0 1 (2.7%) 0 1 (4.2%) 0 Amylaseincreased 1(25%) 0 0 0 0 0 Anemia 0 1 (2.7%) 0 0 0 0 Atrial fibrillation0 1 (2.7%) 0 0 0 0 Cardiomyopathy 0 0 0 0 0 1(33%) GGT increased 0 0 1(2.7%) 0 0 0 Hyperkalemia 0 1 (2.7%) 0 0 0 0 Hypertriglyceridemia 0 1(2.7%) 0 0 0 0 Lacunar infarction 0 0 0 0 1 (4.2%) 0

Safety Profile and Response Rates

TABLE 1 Safety Profile (Incidence Rate ≥ 5%) Any grade Grade 3-4, TRAEsn (%) n (%) Overall 67 (96%) 44 (63%) Hematological 36 (51%) 36 (51%)Thrombocytopenia 25 (50%) 35 (50%) Leucocytopenia 12 (17%) 12 (17%)Anaemia  6 (8.6%)  5 (7.1%) Non-hematological 67 (96%) 11 (16%) Skinhyperpigmentation 17 (24%)  0 Pyrexia 13 (19%)  4 (5.7%) Myalgia 11(16%)  0 Rash  9 (13%)  0 Skin mass  6 (8.6%)  0 Hypertriglyceridaemia36 (51%)  2 (2.9%) Hypocalcaemia 25 (36%)  0 Hyperbilirubinemia 20 (29%) 0 CPK increased 18 (26%)  1 (1.4%) AST increased 17 (24%)  0 ALTincreased 15 (21%)  0 Hypokalaemia 12 (17%)  0 GGT increased 10 (14%)  0Hyperglycaemia  5 (7.1%)  0 Hyponatraemia  5 (7.1%)  0 Creatinineincreased  4 (5.7%)  0

TABLE 2 Response Rates T315I Without T315I All mutation mutationHematological response-n(%) Complete 65/70 40/45 25/25 hematological(93%) (89%) (100%) response-n(%) Cytogenetic response-n(%) Major 24/4717/29  7/18 cytogenetic (51%) (59%) (39%) response-n (%) Complete 14/4713/29  1/18 cytogenetic (30%) (45%) (6%) response-n (%) Molecularresponse Major  6/48  6/30  0/18 molecular (13%) (20%) response-n (%)

Results Analysis

70 patients (CP n=58 and AP n=12) enrolled received ≥1 cycle of thecompound of formula (I-A) treatment, only 1 patient withdrew from thestudy due to disease progression. Median age was 39 (range: 23-59)years. Median interval from CML diagnosis to starting the compound offormula (I-A)-treatment was 6.1 (1.1-14.7) years. Sixty-one (87%)patients received ≥2 prior lines of TKI-therapy. Fifty-three (76%)patients had BCR-ABL mutations and 45 (64%) had T315I mutation atbaseline.

With a median follow-up of 4.1 (1.0-21.2) cycles, the compound offormula (I-A) treatment was well-tolerated in all dose cohorts otherthan the 60 mg cohort. In all patients, 67 (96%) patients experienced ≥1treatment related adverse events (TRAEs) and 44 (63%) experiencedTRAE(s) of grade 3-4 (Table 1). There was no patient withdrawal from thestudy because of TRAEs. Two out of 3 patients in the 60 mg cohortexperienced dose-limiting toxicity (DLT) and the compound of formula(I-A) treatment at 50 mg QOD was considered as maximum toxic dosage(MTD).

The anti-leukemic activities of the compound of formula (I-A) treatmentwere observed in this study. Sixty-five (93%) patients including 58(100%) CP and 7 (58%) AP patients achieved CHR at the dose of 2 mg to 60mg within 3 cycles. In 47 evaluable patients receiving the compound offormula (I-A)-treatment ≥3 cycles, 24 (51%) achieved MCyR including 21(54%) CP and 3 (38%) AP patients at the dose of 12 mg to 50 mg, and 14(30%) patients achieved CCyR including 12 (31%) CP and 2 (25%) APpatients. Total 6 (15%) CP patients achieved MMR. More than 65% of thepatients achieved MCyR or MMR at the end of cycle The compound offormula (I-A) was highly active in patients with or without T315Imutation at baseline (Table 2).

Following oral administration of the compound of formula (I-A) treatmentat doses ranging from 1 mg to 60 mg, the peak concentration was reachedat 2-8 hrs. The elimination appeared to be linear with a mean terminalT_(1/2) of 15.3-36.5 hrs on Day 1, 18.8-42.5 hours on Day 27,respectively (the window period of the observation time is 48 hrs). Theratios for AUC0-t and Cmax of the compound of formula (I-A) treatment onDay 27 versus Day 1 ranged from 1.03 to 2.12 and from 0.78 to 1.93,respectively. Thus, the compound of formula (I-A) exhibited anapproximately dose proportional increase in Cmax and AUC0-t followingsingle or multiple oral administration dose ranging from 1 to 60 mg. PDstudy results demonstrated that reduction of CRKL phosphorylation wasschedule and dose-dependent, ≥50% reduction was observed at dosesranging 12 mg-60 mg.

FIGS. 1A and 1B illustrate the efficacy (CHR n %) of the compound offormula (I-A) in a phase 1 study.

-   -   Wherein, FIG. 1A shows that 98% of the total 82 CML-CP patients        achieved CHR, including 50 patients of T315I⁺ (96% achieved CHR)        and 32 patients of T315I⁻ (100% achieved CHR).    -   FIG. 1B shows that 86% of the total 14 CML-AP patients achieved        CHR, including 11 patients of T315I⁺ (82% achieved CHR) and 3        patients of T315I⁻ (100% achieved CHR).

FIGS. 2A and 2B illustrate the efficacy (MCyR n %) of the compound offormula (I-A) in a phase 1 study.

-   -   Wherein, FIG. 2A shows that 67% of the total 66 CML-CP patients        achieved MCyR, wherein 14% of them achieved PCyR, and 53%        achieved CCyR. 78% of the 42 patients of T315I⁺ achieved MCyR        (of which 4% achieved PCyR, and 74% achieved CCyR); 46% of the        24 patients of T315I⁻ achieved MCyRa (wherein 29% achieved PCyR,        and 17% achieved CCyR).    -   FIG. 2B shows that 25% of the total 12 CML-AP patients achieved        MCyR. Wherein, 33% of the 9 patients of T315I⁺ achieved CCyR; 3        patients of T315I⁻ had a response rate of 0.

FIGS. 3A, 3B and 3C illustrate the efficacy (MCyR n %) of the compoundof formula (I-A) with specific doses (CP) in a phase 1 study.

-   -   Wherein, FIG. 3A shows that, when a dose of 30 mg is        administered, 50% of the total 14 CML-CP patients achieved MCyR        (wherein 14% achieved PCyR and 36% achieved CCyR). Wherein 71%        of the 7 patients of T315I⁺ achieved MCyR (wherein 14% achieved        PCyR and 57% achieved CCyR); 28% of the 7 patients of T315I⁻        achieved MCyR (wherein 14% achieved PCyR and 14% achieved CCyR).    -   FIG. 3B shows that, when a dose of 40 mg is administered, 54% of        the total 18 CML-CP patients achieved MCyR (wherein 16% achieved        PCyR and 38% achieved CCyR). Wherein 60% of the 10 patients of        T315I⁺ achieved MCyR (all of the 60% achieved CCyR); 50% of the        8 patients of T315I⁻ achieved MCyR (wherein 38% achieved PCyR        and 13% achieved CCyR).    -   FIG. 3C shows that, when a dose of 50 mg is administered, 83% of        the total 12 CML-CP patients achieved MCyR (wherein 25% achieved        PCyR and 58% achieved CCyR). Wherein 100% of the 8 patients of        T315I⁺ achieved MCyR (of which 75% achieved CCyR, and 25%        achieved PCyR); 50% of 4 the patients of T315I⁻ achieved MCyR        (wherein 25% achieved PCyR and 25% achieved CCyR).

FIGS. 4A and 4B illustrate MMR (MCyR n %) of the compound of formula(I-A) in a phase 1 study.

-   -   Wherein FIG. 4A shows that 31% of the total 67 CML-CP patients        achieved MMR, wherein 47% of the 43 patients of T315I⁺ achieved        MMR; 4% of the 24 patients of T315I⁻ achieved MMR.    -   FIG. 4B shows that 8% of the total 12 CML-AP patients achieved        MMR. Wherein, 11% of the 9 patients of T315I⁺ achieved MMR; 0%        of the 3 patients of T315I⁻ achieved MMR.

FIGS. 5A and 5B illustrate the plasma concentration-time profiles of thecompound of formula (I-A) in a phase 1 study.

CONCLUSIONS

As shown in FIGS. 1-5 and Tables 1-2, the preliminary results of thephase 1 clinical study showed that the compound of formula (I-A), anovel 3^(rd)-generation TKI, is safe and highly active in treatment ofthe TKI-resistant patients with CML-CP and CML-AP, with or without T315Imutation.

Example 2

Further Efficacy and Safety Results of Phase 1 Study of the compound offormula (I-A) in Patients with Resistant Chronic Myeloid Leukemia

The compound of formula (I-A) is designed for treatment of patients withchronic myeloid leukemia (CML) resistant to current TKI-therapiesincluding those with T315I mutation. This experiment is focus on theefficacy and safety assessment of the compound of formula (I-A) in arelatively long term.

Methods:

An open-label, 3+3 dose escalation, phase 1 trial of the compound offormula (I-A) design to determine maximum tolerated dose (MTD) andidentify dose-limiting toxicities (DLTs) in patients with chronic phase(CP/CML-CP) or accelerated phase (AP/CML-AP) CML resistant to orintolerant of ≥2 prior TKIs or patients with BCR-ABL T315I M after ≥1prior TKI is ongoing. The compound of formula (I-A) was administeredonce every other day (QOD) in 28-day cycles at 11 dose cohorts rangingfrom 1 mg to 60 mg. The eligible patients received continuous treatmentuntil disease progression or unacceptable toxicity, consent withdrawal,or death. The primary efficacy endpoints were major cytogenetic response(MCyR) for CP and complete hematological response (CHR) for AP. MCyRincludes partial cytogenetic response (PCyR) and complete cytogeneticresponse (CCyR). Blood samples were collected at various time points onDay 1-2 and Day 27-28 during cycle 1 for PK analyses.

Results:

Total 101 patients including 87 CML-CP and 14 CML-AP, wherein 71 (70.3%)are male patients, had received the compound of formula (I-A) as asingle agent QOD doses. A total of 62 (61.4%) patients with T315Imutation were included. Median duration of follow-up was 11.2 m (range,1.2-30.6 m). Median age was 40 years (range 20-64y). Median intervalfrom CML diagnosis to starting the compound of formula (I-A) treatmentwas 5.83 years (range 0.3-15.2y). Most Patients (98%) had baseline ECOGstatus 0-1. Patients were heavily pretreated, 83 (83.8%) patientsreceived ≥2 prior lines of TKI-therapy. Two out of 3 patients at 60 mgcohort experienced DLT, and 50 mg QOD was considered as MTD. After MTDdetermined, dose expansion was implemented in the dose levels of 30 mg,40 mg and 50 mg QOD. A total of 56 patients were included in expansionpart.

The compound of formula (I-A) was well-tolerated in all dose cohortswith an exception of 60 mg cohort. In all patients, 101 (100%) patientsexperienced ≥1 treatment related adverse events (TRAEs), the mostfrequent TRAEs were reported as grade 1 or grade 2. The most commongrade 3/4 TRAEs were hematological AEs, including thrombocytopenia(49.5%). The incidences of AEs tended to be dose-dependent. No death andno CTCAE grade 5 events have occurred on study. The incidence of commonTRAEs (≥10%) are shown in Table 3.

TABLE 3 Summary tabulation: Treatment Related Adverse Events Thecompound of formula (I-A) Any Grade Grade 3, 4 SAE Treated Population101 101 101 Preferred Term, n(%) Non-hematological AEs Skin pigmentation 79 (78.2%)  0  0 Hypertriglyceridaemia  55 (54.5%)  8 (7.9%)  0 ASTelevation  37 (36.6%)  3 (3.0%)  0 Proteinuria  35 (34.7%)  5 (5.0%)  0ALT elevation  34 (33.7%)  2 (2.0%)  0 Bilirubin elevation  34 (33.7%) 1 (1.0%)  0 Hypocalcaemia  34 (33.7%)  0  0 GGT elevation  24 (23.8%) 0  0 Hyponatraemia  23 (22.8%)  0  0 Hyperglycaemia  21 (20.8%)  0  0Myalgia  21 (20.8%)  0  0 CPK elevation  20 (19.8%)  2 (2.0%)  0Hypokalaemia  20 (19.8%)  0  0 Pyrexia  18 (17.8%)   7 (6.9%)  1 (1.0%)Rash  15 (14.9%   2 (2.0%)  0 Skin mass  10 (9.9%)   1 (1.0%)  0Hematological AEs Thrombocytopenia  76 (75.2%)  50 (49.5%)  6 (5.9%)Anemia  25 (24.8%)  12 (11.9%)  2 (2.0%) leukopenia  21 (20.8%)  20(19.8%)  0

The compound of formula (I-A) showed the potent anti-leukemic activitiesat doses ≥12 mg QOD. In sixty-eight (67%) evaluable patients, thecompound of formula (I-A) showed potent anti-leukemic activities in CMLpatients. In the 68 evaluable patients with non-CHR at baseline, 63(92.6%) achieved CHR including 52 out of 55 (94.5%) CP patients and 11out of 13 (84.6%) AP patients, respectively. In the 95 evaluablepatients with non-CCyR at baseline, 56 out of 81 (69.1%) CP patientsachieved MCyR including 49 (60.5%) CCyR; and 6 out of 14 (42.9%) APpatients, achieved MCyR including 5 (35.7%) CCyR, respectively. In the100 evaluable patients, 32 out of 86 (37.2%) CP patients and 5 out of 14(35.7%) AP patients achieved MMR, respectively. The compound of formula(I-A) showed highly efficacious in the patients with T315I mutation(Table 4, FIG. 6A, 6B). The response rate and the depth of responsetended to be time dependent (FIG. 7A).

TABLE 4 Phase I Trial in CML: Efficacy Summary CP-CML PatientsAP-CMLPatients With Without With Without Variable All Patients T315Imut. T315I mut. All Patients T315I mut. T315I mut. Population 87 81 3613 11 3 Hematological response

-n( %) No. of evaluable subject-n 55 33 22 13 10 3 Completehematological 52 (94.5%) 32 (97.0%) 20 (90.9%) 11 (84.6%) 8 (80.0%) 3(100.0%) response-n(%) Cytogenetic response

-n(%) No. of evaluable subject-n 81 48 33 14 11 3 Major cytogenetic 56(69.1%) 30 (81.3%) 17 (51.5%) 6 (42.9%) 6 (54.5%) 0 response-n(%)Complete cytogenetic 49 (60.5%) 37 (77.1%) 12 (36.4%) 5 (35.7%) 5(45.5%) 0 response-n(%) Molecular response

No. of evaluated subject-n 86 51 35 14 11 3 Major/Complete molecular 32(37.2%) 27 (52.9%) 5 (14.3%) 5 (35.7%) 5 (45.5%) 0 response-n(%)Complete molecular 18 (20.9%) 13 (29.4%) 3 (8.6%) 2 (14.3%) 2 (18.2%) 0response-n(%) ¹ Only the subjects who have at least one hematologicalassessment post study treatment are included. ² Only the subjects whohave at least one cytogenetic assessment post study treatment isincluded. ³ Only the subjects who have at least one molecular assessmentpost study treatment is included.

indicates data missing or illegible when filed

9 patients (6 CP, 3 AP) had withdrawal from the study, includingprogressive disease (n=5, 2 CP and 3 AP), intolerant AEs (n=2), consentwithdraw (n=1), and secondary breast cancer (n=1). The progression freesurvival (PFS) rate at 18-month was 94% in the CP patients and 61% inthe AP patients (FIG. 7B).

Following a single oral administration of the compound of formula (I-A)at doses from 1-60 mg, the peak concentration of the compound of formula(I-A) was reached between 1-12 h on Day 1, with median Tmax ranging from4-8 h. The elimination appeared to be linear with a mean terminalT_(1/2) of 17.5 to 36.5 h on Day 1. Peak concentration of the compoundof formula (I-A) were observed at 1˜12 h on Day 27, With median Tmaxranging from 4-6 h. The mean terminal T_(1/2) ranged from 19.8 to 42.5 hon Day 27 (both of the watching time window is 48 hrs). The mean ratiosAUC 0-48 h and Cmax of the compound of formula (I-A) on Day 27 to thaton Day 1 ranged from 1.15 to 1.98 and from 0.91 to 1.66, respectively,suggesting moderate accumulation with once every other day dosingregimen. Reduction of CRKL phosphorylation in PBMCs, a biomarker ofBCR-ABL inhibition, has shown to be dose and time dependent in 53evaluable patients treated with the compound of formula (I-A).

Conclusions: the compound of formula (I-A) exhibits significant anddurable antitumor activity, it is well tolerated in the patients withTKI-resistant CML, including those patients with T315I mutation. Theprogression free survival (PFS) rate at 18-month was 94% in the CPpatients and 61% in the AP patients.

In preclinical in vivo studies, compound of the formula (I-A) inducedcomplete regression of subcutaneous tumors in a human CML xenograftmodel and an isogenic model derived from murine Ba/F3 cells expressingBCR-ABL^(WT) or BCR-ABL^(T315I) mutants, and significantly improved thesurvival rate of isogenic leukemia mice carrying Ba/F3 cells havingBCR-ABL^(WT) or BCR-ABL^(T315I). For a mouse model of Ba/F3 tumorcarrying BCR-ABL^(T315I), the (I-A) compound was administered orallyonce every two days (q2d), or once every three days (q3d), and imatinibwas administered once a day (qd) as a control, the results were shown inFIG. 8: compound of the formula (I-A) significantly prolonged thesurvival of BCR-ABL^(T315I) expressing Ba/F3 tumor bearing mice in adose-dependent manner

Example 3

In this experiment, BCR-ABL complex mutation cells were used todetermine the inhibitory effect of the compound of the formula (I-A) andPonatinib on the proliferation of BCR-ABL complex mutation cells. Theexperiment proved that the compound of the formula (I-A) was a potentialeffective medicament capable of overcoming the drug resistance of thePonatinib. Ba/F3 cells stably expressing BCR-ABL (F359V, H396R, E255K,Y253H, T315I, F317L) mutations were provided by the Institute of Lifeand Health, Guangzhou Academy of Sciences.

1. The mutated Ba/F3 cell line stably expressing BCR-ABL (E255V, T315M,Y253H/E255V, Y253H/T315I, Y253H/F359V, T315I/F317L, F317L/F359V)mutation was constructed by electro-transformation method, the Ba/F3cell line stably expressing the BCR-ABL (E255V/T315I, T315I/F359V)mutation was constructed by lentivirus infection method. Cell genesequencing results confirmed that the BCR-ABL mutant gene was integratedinto the genome of Ba/F3 cells. Western blotting results show that theexpression of BCR-ABL protein in these cell lines confirmed the validityof the constructed Ba/F3 stably transfected cell line. The cells werecultured in RPMI 1640 (Base media, Cat #L210KJ) medium containing 10%FBS (GIBCO, Cat #10099-41) and 1% Penicillin-Streptomycin doubleantibodies (Basal Media, Cat #5110JV).

2. Effect of drugs on the proliferation of BaF3 cells stably transfectedwith BCR-ABL single mutation or complex mutation by WST assay

The solution of the sample (the compound of the formula (I-A) orPonatinib) to be tested with a 9-dose concentration obtained by serialdilution was added proportionally at 100 μl/well in a 96-well cultureplate. The dilution was used as a cell blank control (excluding thesample to be tested, which was added to the cells). In addition, anegative control (excluding the sample to be tested and cells) wasprepared. In addition to the negative control wells, 100 μl of completemedium cell suspension was added to each well. The dilution was added tothe negative control wells at 100 μl/well. 3 repeated wells were set inthe experiment. Cells were incubated for 72 hours at 37° C. in a CO₂incubator. 20 μl of CCK-8 detection solution (Shanghai Life iLab BiotechCo., LTD, Cat #D3100L4057) was added to each well, incubating at 37° C.for 2 hours in a CO₂ incubator. The OD value was measured at A 450 nm bya microplate reader.

The percentage of cell viability was calculated using the followingformula:

(O.D. test well−O.D. negative control well)/(O.D. cell control well−O.D.negative control well)×100

The IC₅₀ was calculated using a non-linear regression data analysismethod of Graphpad Prism 6.0 software (Golden software, Golden, Colo.,USA).

3. Results

The compound of the formula (I-A) has better anti-proliferation effecton Ba/F3 cells with complex mutations of BCR-ABL^(E255V/T315I),BCR-ABL^(Y253H/E255V), BCR-ABL^(T315M), BCR-ABL^(Y253H/T315I),BCR-ABL^(Y253H/F359V), BCR-ABL^(T315I/F317L) than Ponatinib, the IC₅₀value of the compound of the formula (I-A) was 2-9 folds lower than theIC₅₀ value of Ponatinib (Table 5). The results suggest that the compoundof the formula (I-A) is a potential candidate drug for overcoming thedrug resistance of Ponatinib caused by BCR-ABL complex mutation.

TABLE 5 Antiproliferative effect of the compound of the formula (I-A) onBa/F3 cells with a single or complex mutation of BCR-ABL Ba/F3containing Antiproliferation Effect (IC₅₀, nM) BCR-ABL the compound ofMutation Region mutation Imatinib Nilotinib Dasatinib Ponatinib theformula (I-A) Wild Type Wild Type  565 ± 656 31 ± 4  10 ± 3 11 6 ± 3Substrate F359V 9626 ± 481 4643 ± 2260  2161 ± 1090 2173 ± 1481 281 ±187 Binding Region A-Ring H396R  9179 ± 1303 4665 ± 799   1641 ± 11802035 ± 1024 274 ± 82  P-Ring E255K 8222 ± 484 648 ± 395 14 ± 1 49 ± 4 22 ± 13 Y253H  8936 ± 1.774 497 ± 122 11 ± 2 37 ± 1  7 ± 1 E255V 4030 ±980 493 ± 163 18 ± 7 42.5 ± 17  13.5 ± 1   Housekeeping T315I 12620 ±28  3425 ± 650  2525 ± 322 33 ± 11 24 ± 10 Gene Hinge F317L 526 ± 56 89± 8  11 ± 1 7 ± 1 8 ± 3 Region Complex E255V/T315I >10000 6467 ± 4431 3571 ± 1385 244 ± 125 26 ± 11 Mutation T315M >10000 >10000  14937 ±10597 1987 ± 1414 217 ± 131 Y253H/E255V >10000 7026 ± 2183 231 ± 92 772± 220 122 T315I/F359V >10000 1944 ± 149  3472 ± 711 17 ± 2  7 ± 1F317L/F359V  3455   354 25 14  12 T315I/F317L  6576 ± 4453 6060 ± 4382 3646 ± 2768 670 ± 405 114 ± 9  Y253H/F359V >10000 9642 ± 591   89 ± 40317 ± 25  100 ± 81  Y253H/T315I >10000 >10000 4767 ± 935 978 ± 57  111 ±1 

We claim:
 1. A method of treating cancer in a patient, comprisingadministering to the patient a therapeutically effective amount of acompound of formula (I):

or a pharmaceutically acceptable salt thereof, wherein R₁ is hydrogen,C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₁₋₄ alkyloxy, or phenyl; and R₂ ishydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or halogen.
 2. The method ofclaim 1, wherein the cancer is hematological malignancy.
 3. The methodof claim 2, wherein the hematological malignancy is leukemia or chronicmyeloid leukemia.
 4. (canceled)
 5. The method of claim 3, wherein themethod is in the treatment of the patient with chronic myeloid leukemiaresistant to current tyrosine kinase inhibitor therapies.
 6. The methodof claim 5, wherein the patient with chronic myeloid leukemia resistantto the current tyrosine kinase inhibitor therapies is caused by BCR-ABLmutations, where the BCR-ABL mutation is T315I, E255K/V, G250E, H396P,M351T, Q252H, Y253F/H, or BCR-ABL^(WT) mutations. 7.-8. (canceled) 9.The method of claim 5, wherein the patient with chronic myeloid leukemiaresistant to the current tyrosine kinase inhibitor therapies is causedby BCR-ABL complex mutation, preferably, wherein the BCR-ABL complexmutation is BCR-ABL^(E255V/T315I), BCR-ABL^(Y253H/E255V),BCR-ABL^(T315M), BCR-ABL^(Y253H/T315I), BCR-ABL^(Y253H/F359V), orBCR-ABL^(T315I/F317L), or one or more of the above-said types ofmutations, and the tyrosine kinase inhibitor is Ponatinib. 10.-11.(canceled)
 12. The method of claim 1, wherein the compound of formula(I), or pharmaceutically acceptable salt thereof is administered orallyevery one, two, or three days during a treatment cycle, wherein saidtreatment cycle is 20-40 days, 25-35 days, or 28-day treatment cycle.13. The method of claim 1, wherein the therapeutically effective amountis from 0.5 mg to 100 mg, from 1 mg to 80 mg, or from 1 mg to 60 mg. 14.The method of claim 13, wherein the therapeutically effective amount isabout 1 mg, about 2 mg, about 4 mg, about 8 mg, about 12 mg, about 20mg, about 30 mg, about 40 mg, about 45 mg, about 50 mg or about 60 mg.15. The method of claim 1, wherein R₁ is selected from the groupconsisting of hydrogen, methyl, cyclopropyl, phenyl, and methoxy; and R₂is selected from the group consisting of hydrogen, methyl, ethyl,cyclopropyl, fluorine, chlorine, or bromine.
 16. The method of claim 1,wherein the compound of formula (I) is a compound of formula (I-A):

or a pharmaceutically acceptable salt thereof.
 17. (canceled)
 18. Amethod of inhibiting BCR-ABL mutants, comprising contacting a compoundof formula (I):

or a pharmaceutically acceptable salt thereof with BCR-ABL mutants,wherein R₁ is hydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₁₋₄ alkyloxy, orphenyl; and R₂ is hydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or halogen. 19.The method of claim 18, wherein the BCR-ABL mutants is T315I, E255K/V,G250E, H396P, M351T, Q252H, Y253F/H, or BCR-ABL^(WT)
 20. (canceled) 21.The method of claim 18, wherein the compound of formula (I) is acompound of formula (I-A):

or a pharmaceutically acceptable salt thereof.
 22. (canceled)
 23. Themethod of claim 18, wherein the inhibition is in a patient with chronicmyeloid leukemia resistant to current tyrosine kinase inhibitortherapies.
 24. The method of claim 23, wherein the patient with chronicmyeloid leukemia resistant to the current tyrosine kinase inhibitortherapies is caused by BCR-ABL mutations, wherein the BCR-ABL mutationis T315I, E255K/V, G250E, H396P, M351T, Q252H, Y253F/H, or BCR-ABL^(WT)mutations. 25.-26. (canceled)
 27. The method of claim 23, wherein thepatient with chronic myeloid leukemia resistant to the current tyrosinekinase inhibitor therapies is caused by BCR-ABL complex mutation,wherein the BCR-ABL complex mutation is BCR-ABL^(E255V/T315I),BCR-ABL^(Y253H/E255V), BCR-ABL^(T315M), BCR-ABL^(Y253H/T315I),BCR-ABL^(Y253H/F359V), or BCR-ABL^(T315I/F317L), or one or more of theabove-said types of mutations, and the tyrosine kinase inhibitor isPonatinib.
 28. (canceled)
 29. The method of claim 23, wherein the methodcomprising orally administering to the patient a therapeuticallyeffective amount of a compound of formula (I-A):

or a pharmaceutically acceptable salt thereof.
 30. The method of claim29, wherein the compound of formula (I), or pharmaceutically acceptablesalt thereof is administered every one, two, or three days during atreatment cycle, wherein said treatment cycle is 20-40 days, 25-35 days,or 28-day treatment cycle.
 31. The method of claim 29, wherein thetherapeutically effective amount is from 0.5 mg to 100 mg, from 1 mg to80 mg, or from 1 mg to 60 mg.
 32. The method of claim 31, wherein thetherapeutically effective amount is about 1 mg, about 2 mg, about 4 mg,about 8 mg, about 12 mg, about 20 mg, about 30 mg, about 40 mg, about 45mg, about 50 mg, or about 60 mg.
 33. A pharmaceutical composition forinhibiting BCR-ABL mutants or for treating hematological malignancy,including chronic myelogenous leukemia, comprising the compound offormula (I) or formula (I-A)

or a pharmaceutically acceptable salt thereof, wherein R₁ is hydrogen,C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₁₋₄ alkyloxy, or phenyl; and R₂ ishydrogen, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, or halogen. 34.-35. (canceled)36. The method of claim 16, wherein the compound of formula (I-A), orpharmaceutically acceptable salt thereof is administered once everyother day (QOD) during a 28-day treatment cycle in an amount at about 1mg, about 2 mg, about 4 mg, about 8 mg, about 12 mg or about 20 mg. 37.(canceled)